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A Vinblastine Fluorescent Probe For Pregnane X Receptor In A Time-resolved Fluorescence Resonance Energy Transfer Assay | 18686
Journal of Analytical & Bioanalytical Techniques
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The pregnane X receptor (PXR) regulates the metabolism and excretion of xenobiotics and endobiotics by regulating the
expression of drug-metabolizing enzymes and transporters. The unique structure of PXR allows the binding of many
drugs and drug leads to it, possibly causing undesired drug-drug interactions. Therefore, it is crucial to evaluate whether lead
compounds bind to PXR. Fluorescence-based assays are preferred because of their sensitivity and nonradioactive nature. One
fluorescent PXR probe is currently commercially available; however, because its chemical structure is not publicly disclosed,
it is not optimal for studying ligand-PXR interactions. Here we report the characterization of BODIPY FL-vinblastine, as a
high-affinity ligand for human PXR with a
value of 673 nM. We provide evidence that BODIPY FL-vinblastine is a unique
chemical entity different from either vinblastine or the fluorophore BODIPY FL in its function as a high-affinity human PXR
ligand. We describe a BODIPY FL-vinblastine-based human PXR time-resolved fluorescence resonance energy transfer assay,
which was used to successfully test a panel of human PXR ligands. The BODIPY FL-vinblastine-based biochemical assay is
suitable for high-throughput screening to evaluate whether lead compounds bind to PXR.
Wenwei Lin has completed his PhD at the University of Tennessee, Health Science Center and postdoctoral studies at the St. Jude Children?s Research Hospital. He is currently a high throughput screening specialist in the High Throughput Screening Center, St. Jude Children?s Research Hospital. He has authored or co- authored more than 20 papers in reputable journals.
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